STANDARDISATION AND HPTLC METHOD DEVELOPMENT OF MARKETED AYURVEDIC FORMULATION – BALARISHTA

Objective: The present study aims to standardize four marketed brands of Balarishta, an Ayurvedic formulation viz. Baidyanath-Balarishta (BB), Dabur-Balarishta (DB), Zandu-Balarishta (ZB) and Nagarjuna-Balarishta (NB) with respect to their physicochemical (organoleptic properties, pH, specific gravity, total solid content, ethanol content, reducing and non-reducing sugar content), phytochemical and microbial parameters (total bacterial count, total fungal count and test for specific pathogens viz. P. aeruginosa, E. coli and S. aureus). It also aims to develop and validate a new highperformance thin layer chromatography (HPTLC) method for simultaneous determination of three major phytoconstituents present in Balarishta viz. withaferin A, gallic acid and ephedrine.Methods: ‘Protocol for testing Ayurveda, Siddha and Unani medicines' was used as a reference for conducting standardization experiments. HPTLC method was developed on Camag Linomat-5 using silica gel 60 GF254 as the stationary phase and Toluene: Chloroform: n-propanol: Ethanol: Formic acid (6: 3: 1: 2: 1, v/v/v/v/v) as the mobile phase. The analytical method validation studies were performed as per International Conference on Harmonization-Quality (ICH-Q2 (R1)) guidelines.Results: The results of standardisation tests obtained were compared with specifications mentioned in ‘Ayurvedic Pharmacopoeia of India 2008 Volume 2, Part 2' and a comparative data of each Balarishta formulation was generated for all the quality control parameters performed. A new, accurate, precise and robust HPTLC method was successfully developed with Retardation factor (Rf) of 0.17±0.02, 0.35±0.01 and 0.54±0.02 for ephedrine, gallic acid and withaferin A respectively.Conclusion: The results of this research work will serve as a valuable quality tool for routine quality control analysis of Balarishta formulations.Keywords: Balarishta, Standardisation, Withaferin A, Gallic acid, Ephedrine and HPTLC

BIOACTIVITY GUIDED FRACTIONATION AND ANTI-INFLAMMATORY ACTIVITY OF ACACIA NILOTICA PODS

Objective: The present study aims at screening the methanol extract and the tannin fraction of the pods of Acacia nilotica for anti-inflammatory activity. Methods: The methanol extract and its tannin fraction were evaluated for acute toxicity using rats as per the Organization for Economic Co-operation and Development (OECD) guidelines. They were found to be safe up to a dose of 2000 mg/kg. The anti-inflammatory activity of the extract and its fraction was evaluated using the carrageenan-induced rat paw edema method for determining the acute phase of inflammation. The activity against sub-acute inflammation was evaluated using the cotton pellet granuloma pouch method. Diclofenac sodium was used as the reference drug.Results: The results of pharmacological screening indicated the methanol extract to possess statistically significant anti-inflammatory activity at 100 mg/kg dose. The tannin fraction of methanol extract was found to be more potent than the methanol extract with maximum response at 25 mg/kg dose.Conclusion: The methanol extract and tannin fraction of pods of Acacia nilotica possess significant anti-inflammatory activity the latter one being more potent.Â

Theoretical study of pressure dependence of superconducting state parameters of some metals using pseudopotential approach

In the present theoretical study, we have calculated superconducting state parameters (SSPs) viz; electron-phonon coupling strength (λ), Coulomb pseudopotential (µ*), critical temperature (Tc), effective interaction strength (N0V) and isotopic effect parameter ( α) of some polyvalent metals (Pb, Ga, In, Sn and Tl) using well-established structured local pseudopotential due to Fiolhais et al. (1995). The pseudopotential with its individual set of parameters has been found to be good in predicting transition temperature Tc for all the metals. Looking to such success, we have extended the present model for the theoretical study of pressure dependence of transition temperature Tc using Debye- Gruneisen model. Our predicted critical volumes using different approaches are well agreed with each other and also with other reported findings. Thus, the present model is consistent and better than nonlocal norm conserving pseudopotentials because it is found to be transferable without any kind of adjustment of its parameters along with its simplicity and predictivity

TRIM32 regulates mitochondrial mediated ROS levels and sensitizes the oxidative stress induced cell death

Emerging evidence suggests that ubiquitin mediated post translational modification is a critical regulatory process involved in diverse cellular pathways including cell death. During ubiquitination, E3 ligases recognize target proteins and determine the topology of ubiquitin chains. Recruitment of E3 ligases to targets proteins under stress conditions including oxidative stress and their implication in cell death have not been systemically explored. In the present study, we characterized the role of TRIM32 as an E3 ligase in regulation of oxidative stress induced cell death. TRIM32 is ubiquitously expressed in cell lines of different origin and form cytoplasmic speckle like structures that transiently interact with mitochondria under oxidative stress conditions. The ectopic expression of TRIM32 sensitizes cell death induced by oxidative stress whereas TRIM32 knockdown shows a protective effect. The turnover of TRIM32 is enhanced during oxidative stress and its expression induces ROS generation, loss of mitochondrial transmembrane potential and decrease in complex-I activity. The pro-apoptotic effect was rescued by pan-caspase inhibitor or antioxidant treatment. E3 ligase activity of TRIM32 is essential for oxidative stress induced apoptotic cell death. Furthermore, TRIM32 decreases X-linked inhibitor of apoptosis (XIAP) level and overexpression of XIAP rescued cells from TRIM32 mediated oxidative stress and cell death. Overall, the results of this study provide the first evidence supporting the role of TRIM32 in regulating oxidative stress induced cell death, which has implications in numerous pathological conditions including cancer and neurodegeneration

Nano-enabled biosensing systems for intelligent healthcare: towards COVID-19 management

Biosensors are emerging as efficient (sensitive and selective) and affordable analytical diagnostic tools for early-stage disease detection, as required for personalized health wellness management. Low-level detection of a targeted disease biomarker (pM level) has emerged extremely useful to evaluate the progression of disease under therapy. Such collected bioinformatics and its multi-aspects-oriented analytics is in demand to explore the effectiveness of a prescribed treatment, optimize therapy, and correlate biomarker level with disease pathogenesis. Owing to nanotechnology-enabled advancements in sensing unit fabrication, device integration, interfacing, packaging, and sensing performance at point-of-care (POC) has rendered diagnostics according to the requirements of disease management and patient disease profile i.e. in a personalized manner. Efforts are continuously being made to promote the state of art biosensing technology as a next-generation non-invasive disease diagnostics methodology. Keeping this in view, this progressive opinion article describes personalized health care management related analytical tools which can provide access to better health for everyone, with overreaching aim to manage healthy tomorrow timely. Considering accomplishments and predictions, such affordable intelligent diagnostics tools are urgently required to manage COVID-19 pandemic, a life-threatening respiratory infectious disease, where a rapid, selective and sensitive detection of human beta severe acute respiratory system coronavirus (SARS-COoV-2) protein is the key factor

The Abdominal Aortic Aneurysm Statistically Corrected Operative Risk Evaluation (AAA SCORE) for predicting mortality after open and endovascular interventions

BackgroundAccurate adjustment of surgical outcome data for risk is vital in an era of surgeon-level reporting. Current risk prediction models for abdominal aortic aneurysm (AAA) repair are suboptimal. We aimed to develop a reliable risk model for in-hospital mortality after intervention for AAA, using rigorous contemporary statistical techniques to handle missing data.MethodsUsing data collected during a 15-month period in the United Kingdom National Vascular Database, we applied multiple imputation methodology together with stepwise model selection to generate preoperative and perioperative models of in-hospital mortality after AAA repair, using two thirds of the available data. Model performance was then assessed on the remaining third of the data by receiver operating characteristic curve analysis and compared with existing risk prediction models. Model calibration was assessed by Hosmer-Lemeshow analysis.ResultsA total of 8088 AAA repair operations were recorded in the National Vascular Database during the study period, of which 5870 (72.6%) were elective procedures. Both preoperative and perioperative models showed excellent discrimination, with areas under the receiver operating characteristic curve of .89 and .92, respectively. This was significantly better than any of the existing models (area under the receiver operating characteristic curve for best comparator model, .84 and .88; P < .001 and P = .001, respectively). Discrimination remained excellent when only elective procedures were considered. There was no evidence of miscalibration by Hosmer-Lemeshow analysis.ConclusionsWe have developed accurate models to assess risk of in-hospital mortality after AAA repair. These models were carefully developed with rigorous statistical methodology and significantly outperform existing methods for both elective cases and overall AAA mortality. These models will be invaluable for both preoperative patient counseling and accurate risk adjustment of published outcome data

Feeding habits of milk shark, Rhizoprionodon acutus (Ruppell, 1837) in the Gujarat coastal waters of north-eastern Arabian Sea

The feeding habit of milk shark Rhizoprionodon acutus was investigated in 684 specimens collected along Gujarat coast (India) of north-eastern Arabian Sea from January 2013 to December 2014. The length range of females was 32–89.6 cm and males 33.5–89 cm. The shark foraged on diversified prey items which were pooled into four distinct groups i.e., teleosts, crustaceans, mollusks and annelids. Teleosts (Dietary coefficient, %QI = 83.05 and index of relative importance, %IRI = 78.40) were found to be the preferred food items followed by crustaceans (%QI = 16.21; %IRI = 19.78), which formed the secondary food item group. Mollusks (%QI = 0.74; %IRI = 1.69) and annelids (%QI = 0.01; %IRI = 0.14) constituted the accidental or accessory food items. The species, though is a pelagic predator, probably performs vertical movements in search of prey items. The shark also showed some sorts of preference and selectivity for clupeids, engraulids and carangids. Females though showed significantly lower index of relative fullness (IRF) (P ≤ 0.5) and comparatively lower vacuity index and lower mean number of preys per stomach compared to the males, the preference for prey items was not found to be significantly different between the females and males. Juveniles were found to have significantly higher IRF (P ≤ 0.5) and comparatively higher vacuity index than that of the adults, whereas the mean preys per stomach was found to be lower than that of the adults. Moreover, the prey preference was also significantly different between the juveniles and adults. The study provides necessary baseline information about the feeding habits of the shark in the region which will be helpful in understanding the trophodynamics of the species under the influence of overfishing and climate change

Exosome Release Is Modulated by the Mitochondrial-Lysosomal Crosstalk in Parkinson’s Disease Stress Conditions

Parkinson’s disease (PD) is a neurodegenerative disorder characterized by the progressive loss of dopaminergic neurons in the substantia nigra (SN) pars compacta region of the brain. The main pathological hallmark involves cytoplasmic inclusions of α-synuclein and mitochondrial dysfunction, which is observed in other part of the central nervous system other than SN suggesting the spread of pathogenesis to bystander neurons. The inter-neuronal communication through exosomes may play an important role in the spread of the disease; however, the mechanisms are not well elucidated. Mitochondria and its role in inter-organellar crosstalk with multivesicular body (MVB) and lysosome and its role in modulation of exosome release in PD is not well understood. In the current study, we investigated the mitochondria-lysosome crosstalk modulating the exosome release in neuronal and glial cells. We observed that PD stress showed enhanced release of exosomes in dopaminergic neurons and glial cells. The PD stress condition in these cells showed fragmented network and mitochondrial dysfunction which further leads to functional deficit of lysosomes and hence inhibition of autophagy flux. Neuronal and glial cells treated with rapamycin showed enhanced autophagy and inhibited the exosomal release. The results here suggest that maintenance of mitochondrial function is important for the lysosomal function and hence exosomal release which is important for the pathogenesis of PD

TNF-α-induced E3 ligase, TRIM15 inhibits TNF-α-regulated NF-κB pathway by promoting turnover of K63 linked ubiquitination of TAK1

Ubiquitin E3-ligases are recruited at different steps of TNF-α-induced NF-κB activation; however, their role in temporal regulation of the pathway remains elusive. The study systematically identified TRIMs as potential feedback regulators of the TNF-α-induced NF-κB pathway. We further observed that TRIM15 is “late” response TNF-α-induced gene and inhibits the TNF-α-induced NF-κB pathway in several human cell lines. TRIM15 promotes turnover of K63-linked ubiquitin chains in a PRY/SPRY domain-dependent manner. TRIM15 interacts with TAK1 and inhibits its K63-linked ubiquitination, thus NF-κB activity. Further, TRIM15 interacts with TRIM8 and inhibits cytosolic translocation to antagonize TRIM8 modualted NF-κB. TRIM8 and TRIM15 also show functionally inverse correlation in psoriasis condition. In conclusion, TRIM15 is TNF-α-induced late response gene and inhibits TNF-α induced NF-κB pathway hence a feedback modulator to keep the proinflammatory NF-κB pathway under control